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Method for constructing HSAT H-T-S mouse-human chimeric model and application

A technology of HSATH-T-S and a construction method is applied in the field of HSATH-T-S human-mouse chimera model construction, which can solve the problems of reduced supply of human leukocyte antigens, opportunistic infections, loss of function of grafts, etc., and achieves good application prospects, Reproducible and effective results

Inactive Publication Date: 2009-05-27
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, immunosuppressants, T-cell depletion transplantation, and tissue matching are mainly used to prevent and treat transplant rejection, but the side effects such as tumor induction, opportunistic infection, and graft loss of function are increasingly apparent, and the supply of human leukocyte antigen continues to decrease. , seeking new ways to prevent and treat transplant rejection has become the focus of research in the field of transplantation

Method used

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  • Method for constructing HSAT H-T-S mouse-human chimeric model and application
  • Method for constructing HSAT H-T-S mouse-human chimeric model and application
  • Method for constructing HSAT H-T-S mouse-human chimeric model and application

Examples

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Effect test

Embodiment 1

[0041] Example 1 Establishment of HSAT H-T-S human-mouse chimera animal model, the steps are as follows:

[0042] Experimental Materials:

[0043] Animals: 20 CB-17SCID mice (purchased from the Animal Experiment Center of Wuhan University), about 8 weeks old, female, weighing about 20g

[0044] Raising conditions: under sterile conditions, raising temperature 22°C, humidity 55%, 12-hour white / circadian rhythm, giving appropriate antibiotics in drinking water

[0045] Anesthetics: xylazine hydrochloride (0.3 mg, Liaoning Satellite Pharmaceutical Factory), ketamine hydrochloride (1.5 mg, Liaoning Satellite Pharmaceutical Factory), 1×PBS buffer 300 μl

[0046] A construction method of HSATH-T-S human-mouse chimera animal model, comprising the following steps:

[0047] A. Twenty CB-17SCID mice (non-obese diabetic mice, purchased from the Animal Experiment Center of Wuhan University) about 8 weeks old were selected and divided into two groups. Four hours before cell transplantat...

Embodiment 2

[0052] Embodiment 2 A kind of HSAT H-T-S human-mouse chimera model Its application process is as follows: (the detection of HSAT H-T-S human-mouse chimera animal model)

[0053] Experimental materials: After the HSAT H-T-S human-mouse chimera model was established, under sterile conditions, the thymus, spleen, liver and peripheral blood were obtained from the organs of the two groups of SCID mice at 0, 4, 8, and 12 weeks, respectively. Single-cell suspension of lymphocytes was collected. The method for isolating lymphocytes from mouse organs in vitro is as follows: first cut the organs into small pieces, pass through 200-mesh stainless steel wire mesh, and then use Percoll separation medium (Pharmacia, USA) to separate lymphocytes. For detailed steps, please refer to papers such as Mebius RE (Transfer of primitive stem / progenitor bonemarrow cells from LT alpha- / - donors to wild-type hosts: implications for the generation of architectural events in lymphoid B cell domains. JImm...

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Abstract

The invention discloses a method for constructing an HSAT H-T-S human-mouse chimera model and the application thereof. Firstly, a female Nod-SCID mouse in 6-8 weeks is selected; whole body exposure is carried out on the SCID mouse by radiation; secondly, human embryo thymic cells are taken; the cells come from the thymus of the embryo of a person who is voluntary to terminate the pregnancy, and the pregnant week is less than 24 weeks; the quantity of the cells is 1*10<7>, wherein, the quantity proportion of the thymic cells, as well as thymic epithelia and other stroma cells is 1:1, and the specificity of alpha-GalCer-CD1d tetramer is used for eliminating young and mature NKT cells; thirdly, the obtained embryo thymic cells are injected to the thymus of the SCID mouse subcutaneously to carry out dermatoplasty and obtain an HSAT H-T-S human-mouse chimera model. In the method, the human embryo thymic cells are implanted to the thymus of the mouse with severe combined immunodeficiency, and the full skin of human embryo trunk is grafted to the chest cavity of the SCID mouse, thereby reproducing the immune tolerance and rejection reaction of the dermatoplasty reaction in human body. The invention has good repeatability, obvious effect, simple method and good application prospect.

Description

technical field [0001] The present invention relates to the technical fields of cellular immunology and experimental zoology, more specifically to a method for constructing a HSATH-T-S human-mouse chimera model, and also relates to the transplantation immune tolerance of NKT cells reproduced in the HSAT H-T-S human-mouse chimera model. role in acceptance and rejection. Background technique [0002] In recent years, studies on animal experimental models have been reported successively, such as the transplantation of human fetal thymus, fetal liver and fetal bone, and even adult peripheral blood hematopoietic stem cells into patients with severe combined immunodeficiency syndrome (severe combined immunodeficiency syndrome, SCID). In mice, several human-mouse chimeric models such as SCIDhyTHy / Liv, SCID huBone, and SCID HPBL were established. Among them, human peripheral blood lymphocytes (HuPBL) have the advantages of convenient source, T and B lymphocytes have functions in th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61B19/00G09B23/28A61B90/00A61D1/00
Inventor 谭锦泉何玉玲陈朗肖睿璟蒋艳萍王岚
Owner WUHAN UNIV
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